Specific nNOS inhibitor cpd8. (A) Docking model of cpd8 with nNOS protein. (B) cpd8 significantly reduced total NOS activities in human melanoma Lu1205 cells. Spermidine trihydrochloride (ST), 10 μM; cpd8, 1 μM. Results are the mean±SD of three biologic replicates of a representative experiment. *p<0.05 compared to control cells. (C, D) cpd8 inhibited nNOS expression induced by DETA/NO stress (C) and UVB radiation (D). Pretreatment: pretreated with cpd8 for 24 h, followed by UVB radiation; cotreatment: radiated by UVB, then replaced with fresh growth media containing cpd8. Twenty-four hours later, whole cell lysates were collected for immunoblotting analysis. (E, F) Elevated proliferation (E) and invasion potential (F) by DETA/NO stress were also reversed by cpd8 cotreatment. *p<0.05, compared to DETA alone (n=2). Proliferation was detected by MTT colorimetric assay (n=3). To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars